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利用来自高度保守的聚合酶区域的引物,通过聚合酶链反应鉴定未表征的逆转录病毒。

The use of primers from highly conserved pol regions to identify uncharacterized retroviruses by the polymerase chain reaction.

作者信息

Donehower L A, Bohannon R C, Ford R J, Gibbs R A

机构信息

Division of Molecular Virology, Baylor College of Medicine, Houston, Texas 77030.

出版信息

J Virol Methods. 1990 Apr;28(1):33-46. doi: 10.1016/0166-0934(90)90085-t.

Abstract

Two degenerate oligonucleotide primers derived from regions of pol conserved among retroviruses have been synthesized. Polymerase chain reactions utilizing these primers amplify a 135-bp pol fragment in every retrovirus DNA tested to date. The polymerase chain reaction has been linked to a reverse transcriptase step so that a pol-specific DNA fragment can be obtained from a moderate amount of a purified retrovirus or viral RNA. The identity of an unknown retrovirus can be determined by sequencing of the amplified fragment following molecular cloning. This procedure was tested on an unidentified (non-HIV) retrovirus expressed by a B-cell lymphoma line obtained from an AIDS patient. Our PCR assay identified the retrovirus as being highly similar to Mason-Pfizer monkey virus (MPMV) and simian retrovirus 1, which are closely related immunosuppressive type D viruses that cause simian AIDS.

摘要

已经合成了两种源自逆转录病毒中pol保守区域的简并寡核苷酸引物。利用这些引物进行的聚合酶链反应在迄今为止测试的每种逆转录病毒DNA中扩增出一个135bp的pol片段。聚合酶链反应已与逆转录步骤相连接,以便可以从适量的纯化逆转录病毒或病毒RNA中获得pol特异性DNA片段。通过分子克隆后对扩增片段进行测序,可以确定未知逆转录病毒的身份。该程序在从一名艾滋病患者获得的B细胞淋巴瘤系所表达的一种未鉴定(非HIV)逆转录病毒上进行了测试。我们的PCR检测确定该逆转录病毒与梅森 - Pfizer猴病毒(MPMV)和猿猴逆转录病毒1高度相似,它们是密切相关的免疫抑制性D型病毒,可导致猿猴艾滋病。

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